Solid-phase bioconjugation of heterobifunctional adaptors for versatile assembly of bispecific targeting ligands

HongYan Liu, Pavel Zrazhevskiy, Xiaohu Gao

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

High-throughput generation of bispecific molecules promises to expedite the discovery of new molecular therapeutics and guide engineering of novel multifunctional constructs. However, high synthesis complexity and cost have hampered the discovery of bispecific molecules in drug development and biomedical research. Herein we describe a simple solid-phase bioconjugation procedure for preparation of Protein A(G,L)-PEG-Streptavidin heterobifunctional adaptors (with 1:1:1 stoichiometry), which enable self-assembly of unmodified antibodies and biotinylated molecules into bispecific targeting ligands in a versatile mix-and-use manner. Utility of such adaptors is demonstrated by assembly of anti-CD3 and anti-Her2 antibodies into bispecific CD3xHer2 targeting ligands, which efficiently drive T-cell-mediated lysis of Her2-positive cancer cells. In comparison to bioconjugation in solution, the solid-phase procedure described here offers precise stoichiometry control, ease of purification, and high yield of functional conjugates. Simplicity and versatility should prove this methodology instrumental for preparation of bispecific ligands, as well as for high-throughput screening of bispecific combinations, before proceeding to synthesis of lead candidates via recombinant engineering or chemical cross-linking.

Original languageEnglish (US)
Pages (from-to)1511-1516
Number of pages6
JournalBioconjugate Chemistry
Volume25
Issue number8
DOIs
StatePublished - Aug 20 2014

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Ligands
Antibodies
Stoichiometry
Molecules
Throughput
Chemical Engineering
Streptavidin
T-cells
Staphylococcal Protein A
Self assembly
Polyethylene glycols
Purification
Biomedical Research
Anti-Idiotypic Antibodies
Screening
Cells
Proteins
T-Lymphocytes
Costs and Cost Analysis
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology
  • Medicine(all)

Cite this

Solid-phase bioconjugation of heterobifunctional adaptors for versatile assembly of bispecific targeting ligands. / Liu, HongYan; Zrazhevskiy, Pavel; Gao, Xiaohu.

In: Bioconjugate Chemistry, Vol. 25, No. 8, 20.08.2014, p. 1511-1516.

Research output: Contribution to journalArticle

Liu, HongYan ; Zrazhevskiy, Pavel ; Gao, Xiaohu. / Solid-phase bioconjugation of heterobifunctional adaptors for versatile assembly of bispecific targeting ligands. In: Bioconjugate Chemistry. 2014 ; Vol. 25, No. 8. pp. 1511-1516.
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